ERK1/ERK2 Recombinant Polyclonal Antibody (6HCLC)/ERK1 / ERK2重组多克隆抗体（6HCLC）

货号：710318

规格：100 µg

价格：4809

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Phosopeptide corresponding to amino acids 197–209 of human MAPK [pT202/pY204]

物种：人/小鼠

宿主：兔

抗体亚型：IgG

克隆号：6HCLC

荧光染料：其它

类型:	多抗	同型对照：
浓度：	0.5 mg/mL	用法：	1 µL(ChIP)；0.5-2 µg/mL(WB)

产品详细信息This antibody is predicted to react with mouse, rat, non-human primate and rabbit based on sequence homology.Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.靶标信息ERK1 and ERK2 are widely expressed and are involved in the regulation of meiosis, mitosis, and postmitotic functions in differentiated cells. Many different stimuli, including growth factors, cytokines, virus infection, ligands for heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors and transforming agents, activate the ERK1 and ERK2 pathways. When growth factors bind to the receptor tyrosine kinase, Ras interacts with Raf, the serine/threonine protein kinase and activates it as well. Once actived, Raf phosphorylates serine residue in 2 further kinases, MEK1/2, which in turn phosphorylates tyrosine/threonine in extracellular-signal regulated kinase (ERK) 1/2. Upon activation, the ERKs either phosphorylate a number of cytoplasmic targets or migrate to the nucleus, where they phosphorylate and activate a number of transcription factors such as c-Fos and Elk-1.

数据：

ERK1/ERK2 Antibody (710318) in WBWestern blot analysis of p44/42 MAPK (ERK1/2) (pT202/pY204) was performed by loading 20 µg of NIH/3T3 (lane1) and NIH/3T3 treated for 10 minutes with 100 nM/mL of EGF (lane2) cell lysates using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for overnight at 4°C. p44/42 MAPK (ERK1/2) (pT202/pY204) was detected at ~42 kDa using p44/42 MAPK (ERK1/2) (pT202/pY204) Recombinant Rabbit Polyclonal Antibody (Product # 710318) at 0.5-1 µg/mL in 2.5% skim milk at room temperature for 3 hours on a rocking platform. Goat anti-Rabbit IgG - HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106). ERK1/ERK2 Antibody (710318) in ChIPChromatin immunoprecipitation analysis of ERK1+ERK23 was performed using cross-linked chromatin from 1 x 10^6 HCT116 human colon carcinoma cells treated with serum for 0, 15, and 60 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of an ERK1+ERK23 Recombinant Rabbit Polyclonal Antibody (Product # 710318). Chromatin aliquots from ~1 x 10^5 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify -3.2kb upstream of the human FOS gene, or exon-4 of human FOS. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the FOS locus is shown above the data where boxes represent exons (grey boxes = translated regions, white boxes = untranslated regions), the zigzag lines represent introns, and the straight line represents upstream sequence. Regions amplified by FOS primers are represented by black bars. Data courtesy of the Innovators Program.

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